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 EFFECTS OF DIETARY PROBIOTICS ON THE GROWTH AND IMMUNITY OF TOR TAMBROIDES  : A MOLECULAR APPROACH WITH EMPHASIS ON GENE EXPRESSION PROFILES Li Lian Wo ng*, Nguyen Thi Mai, Nursyafiqah Zahidah Azahar, Zulaikha Mat Deris and  Norazimah Y us of !hool of "isheries and Aqua!ulture !ien!es, #ni$ersiti Malaysia Te rengganu %mail& lilian'umt(edu(my Malaysian mahseer  )Tor tambroides has signifi!ant aestheti! and e!onomi! $alue as food and ornamental fish( The natural +o+ulation of T. tambroides is !urrently suffering a dramati! de!line in +o+ulation size due to o$ ere-+loitation and en$ironmental degradation( Due to the redu!ed  +o+ulation size and high market demand, artifi!ial .reeding +rogram of this s+e!ies ha$e .een de$elo+ed for .oth !onser$ation and aqua!ulture +ur+oses( /o0e$er, the gro0th rate of T. tambroides  is highly redu!ed in !a+ti$e .reeding !ondition, and !ould only .e im+ro$ed .y administration of e-+ensi$e high +rotein diet( To redu!e the total !ost of aqua!ulture +rodu!tion,  .enefi!ial dietary su++lements su!h as +ro.ioti!s !ould .e used as +romising alternati $es for $ia.le farming of this s+e!ies( 1ro.ioti!s ha$e .een in!luded in fish diet for numerous .enefits 0hi!h in!lude enhan!ing the gro0th and im+ro$ing disease resistan!e of the !u ltured s+e!ies( 2n this study, 0e set out to determine 0hether addition of !ommer!ial +ro.ioti!s )  Bacillus subtilis in the 34day diet 0ould im+ro$e the gro0th rate and immunity le$els of T. tambroides( Measures of .ody 0eight and standard leng th 0ere +erformed to assess 5u$eniles6 gr o0th( The immunity le$el and gro0th rate 0ere o.ser$ed through !om+arison of mole!ular res+onses follo0ing +ro.ioti!s adminis tration in !ontrol diet and +ro.ioti! su++lemented diet( 2nsulinlike gro0th fa!tor )  Igf  gene 0as !hosen as gro0th marker, 0hile /eatsho!k +rotein 74 )  Hsp70 and 8lutathione +ero-idase ) GPX  0ere sele!ted as immunerelated markers( We o.ser$ed signifi!ant in!rease in the .ody 0eight of 5u$en ile fish treated 0ith +ro.ioti! su++lemented diet, and the gro0th rate im+ro$ement 0as further +ro$ed .y a signifi!ant 9(: fold in!rease in the  Igf gene e-+ression le$el( /o0e$er, there 0as no signifi!ant differen!e in the e-+ression le$els for  .oth Hsp70 and GPX genes .et0een !ontrol and +ro.ioti! treated grou+s( This data 0as resulted from the a.sen!e of !hallenge test 0ith e-ternal stressors su!h as +athogeni! infe!tion, +oor 0ater quality and e-treme tem+erature to indu!e the immunity genes e-+ression( ;$erall, this  +reliminary study suggests a +otent effe!t on the gro0t h of 5u$enile T. tambroides, 0hi!h  +ro$ides +rimary insight into mole!ular entry +ath to in$estigate +ro.ioti! me!hanisms res+onsi.le for gro0th enhan!ement of this s+e!ies( With a++ro+riate stress indu!ers in!or+orated into the e-+erimental !hallenge test in our ne-t study, 0e anti!i+ate to +ro$e the effi!ien!y of dietary +ro.ioti!s in regulating immunerelated genes in T. tambroides(

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EFFECTS OF DIETARY PROBIOTICS ON THE GROWTH AND IMMUNITY OF TOR TAMBROIDES : A MOLECULAR APPROACH WITH EMPHASIS ON GENE EXPRESSION PROFILESLi Lian Wong*, Nguyen Thi Mai, Nursyafiqah Zahidah Azahar, Zulaikha Mat Deris and Norazimah YusofSchool of Fisheries and Aquaculture Sciences, Universiti Malaysia TerengganuEmail: [email protected]

Malaysian mahseer (Tor tambroides) has significant aesthetic and economic value as food and ornamental fish. The natural population of T. tambroides is currently suffering a dramatic decline in population size due to over-exploitation and environmental degradation. Due to the reduced population size and high market demand, artificial breeding program of this species have been developed for both conservation and aquaculture purposes. However, the growth rate of T. tambroides is highly reduced in captive breeding condition, and could only be improved by administration of expensive high protein diet. To reduce the total cost of aquaculture production, beneficial dietary supplements such as probiotics could be used as promising alternatives for viable farming of this species. Probiotics have been included in fish diet for numerous benefits which include enhancing the growth and improving disease resistance of the cultured species. In this study, we set out to determine whether addition of commercial probiotics (Bacillus subtilis) in the 30-day diet would improve the growth rate and immunity levels of T. tambroides. Measures of body weight and standard length were performed to assess juveniles growth. The immunity level and growth rate were observed through comparison of molecular responses following probiotics administration in control diet and probiotic supplemented diet. Insulin-like growth factor (Igf) gene was chosen as growth marker, while Heat-shock protein 70 (Hsp70) and Glutathione peroxidase (GPX) were selected as immune-related markers. We observed significant increase in the body weight of juvenile fish treated with probiotic supplemented diet, and the growth rate improvement was further proved by a significant 1.2 fold increase in the Igf gene expression level. However, there was no significant difference in the expression levels for both Hsp70 and GPX genes between control and probiotic treated groups. This data was resulted from the absence of challenge test with external stressors such as pathogenic infection, poor water quality and extreme temperature to induce the immunity genes expression. Overall, this preliminary study suggests a potent effect on the growth of juvenile T. tambroides, which provides primary insight into molecular entry path to investigate probiotic mechanisms responsible for growth enhancement of this species. With appropriate stress inducers incorporated into the experimental challenge test in our next study, we anticipate to prove the efficiency of dietary probiotics in regulating immune-related genes in T. tambroides.